rabbit α- vglut1  (Synaptic Systems)

Journal: Immunity

Article Title: Targeted complement inhibition at synapses prevents microglial synaptic engulfment and synapse loss in demyelinating disease

doi: 10.1016/j.immuni.2019.12.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit pAb α-VGluT1 (human/marmoset tissue) , Millipore , Cat. #abN1627.

Techniques: Virus, Recombinant, Adjuvant, Electron Microscopy, Cloning, Plasmid Preparation, RNAscope, Multiplex Assay, In Situ, Sequencing, Generated, Software, Imaging

Journal: bioRxiv

Article Title: Maturation of Purkinje cell firing properties relies on granule cell neurogenesis

doi: 10.1101/2020.05.20.106732

Figure Lengend Snippet: A. Schematic of an embryonic brain. Inset is the cerebellar anlage. Atoh1 domain (granule cell precursors, pink), Ptf1a domain (Purkinje cell precursors, green), En1 domain (grey). Orientation is the same for all panels unless otherwise indicated. B. Schematic of a sagittal section of a P14 cerebellum. Purkinje cell=green; granule cell=pink. Cerebellar lobules are labelled with Roman numerals . C. Intersectional labelling of En1;Atoh1 domain with tdTomato (pink) shows no overlap with Purkinje cells (Calbindin; green). D. Whole brain images of control and En1 Cre/+ ;Atoh1 fl/- mice showing abnormal gross morphology. E. Sagittal sections of control and En1 Cre/+ ;Atoh1 fl/- hindbrains stained with cresyl violet to visualize cell nuclei. F. and G. Sagittal and coronal sections of P14 cerebella of control ( F ) and En1 Cre/+ ;Atoh1 fl/- ( G ) mice stained with Calbindin (grey). Arrows indicate Purkinje cells that have migrated into the colliculi. F. and G. are presented at the same magnification. H. Higher magnification images of Calbindin staining in control and En1 Cre/+ ;Atoh1 fl/- mice. Images are representative for N=3 brains for each genotype.

Article Snippet: The following primary antibodies were used for the data described in this manuscript: guinea pig (gp)-α-Calbindin (1:1,000; SySy; #214004); rabbit (rb)-α-gamma-aminobutyric acid receptor α6 (GABARα6; 1:500; Millipore Sigma; #AB5610), rb-α-T-box brain protein 2 (Tbr2; 1:500; Abcam; #AB23345), mouse (ms)-α-Calretinin (1:500; Swant; #6B3); ms-α-Neurofilament Heavy (NFH; 1:1,000; Biolegend; #801701); rb-α-Hyperpolarization Activated Cyclic Nucleotide Gated Potassium Channel 1 (HCN1; 1:500; Alomone Lab; #APC-056); goat (gt)-α-RAR-related orphan receptor alpha (RORα; 1:250; Santa Cruz; #F2510); rb-α-parvalbumin (PV; 1:1,000; Swant; #PV25); rb-α-neurogranin (1:500; Millipore Sigma; #AB5620); ms-α-ZebrinII (1:500; kind gift from Dr. Richard Hawkes, University of Calgary, Calgary, Alberta, Canada); rb-α-Vglut1 (1:500; SySy; #135302); rb-α-Vglut2 (1:500; SySy; #135403).

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Maturation of Purkinje cell firing properties relies on granule cell neurogenesis

doi: 10.1101/2020.05.20.106732

Figure Lengend Snippet: A. Schematic of a control Purkinje cell and an En1 Cre/+ ;Atoh1 fl/- Purkinje cell (based on results from B-H ). MF=Mossy Fiber; CF=Climbing fiber. B. Images of Calbindin (green) and Vglut1 (pink) staining. For En1 Cre/+ ;Atoh1 fl/- mice in B, C, and E c=cerebellum, and d=displaced Purkinje cells. Images are representative for N=3 brains for each genotype. C. Images of Calbindin (green) and Vglut2 (pink) staining. D. Schematic of a control Purkinje cell, the En1 Cre/+ ;Atoh1 fl/- ZebrinII staining pattern (see Supp. Figure 2 ) and WGA-Alexa 555 tracing form the spinal cord. E. Representative images of WGA-Alexa 555+ terminals in the cerebellum. Dotted lines represent the border between ZebrinII-positive (cyan) and -negative region (left four images) or between the cerebellum and colliculi (right images). Black and white image shows the pattern of WGA-Alexa 555 positive terminals. F. Representative images of Golgi-Cox-labelled Purkinje cells in control (top row) and En1 Cre/+ ;Atoh1 fl/- brains (bottom row). G. Sholl analysis for dendritic complexity. H. Purkinje cells in En1 Cre/+ ;Atoh1 fl/- mice have shorter and less branched Purkinje cell dendrites (n=30/N=3 for each genotype, each animal is indicated with a differentially oriented triangle). Linear mixed model with genotype as the fixed effect and mouse number as the random effect. *P<0.001 for both distance from soma and branch number. All images were acquired from the cerebellum of P14 mice.

Article Snippet: The following primary antibodies were used for the data described in this manuscript: guinea pig (gp)-α-Calbindin (1:1,000; SySy; #214004); rabbit (rb)-α-gamma-aminobutyric acid receptor α6 (GABARα6; 1:500; Millipore Sigma; #AB5610), rb-α-T-box brain protein 2 (Tbr2; 1:500; Abcam; #AB23345), mouse (ms)-α-Calretinin (1:500; Swant; #6B3); ms-α-Neurofilament Heavy (NFH; 1:1,000; Biolegend; #801701); rb-α-Hyperpolarization Activated Cyclic Nucleotide Gated Potassium Channel 1 (HCN1; 1:500; Alomone Lab; #APC-056); goat (gt)-α-RAR-related orphan receptor alpha (RORα; 1:250; Santa Cruz; #F2510); rb-α-parvalbumin (PV; 1:1,000; Swant; #PV25); rb-α-neurogranin (1:500; Millipore Sigma; #AB5620); ms-α-ZebrinII (1:500; kind gift from Dr. Richard Hawkes, University of Calgary, Calgary, Alberta, Canada); rb-α-Vglut1 (1:500; SySy; #135302); rb-α-Vglut2 (1:500; SySy; #135403).

Techniques: Control, Staining

Journal: Cell Reports

Article Title: TSPAN5 Enriched Microdomains Provide a Platform for Dendritic Spine Maturation through Neuroligin-1 Clustering

doi: 10.1016/j.celrep.2019.09.051

Figure Lengend Snippet: TSPAN5 Is Expressed in the Postsynaptic Compartment of Hippocampal Pyramidal Neurons (A) Confocal images of rat hippocampal cultured neurons immunolabeled for TSPAN5 at DIV 6, 11, and 18. Scale bar, 20 μm. Insets show higher magnification of regions highlighted in white. n = 3 independent cultures. (B) Confocal images of DIV 18 rat hippocampal cultured neurons immunolabeled for TSPAN5 (green) and the synaptic markers (red) PSD-95, GluA2, GABA A R-β3, Bassoon, VGluT1, and VGAT. Insets show higher magnification of regions highlighted in white. Scale bar, 20 μm. (C) Quantification of percentages of TSPAN5 puncta colocalizing with the different markers analyzed. n = 10 neurons. (D) Synaptosomes purification from adult rat hippocampi and cortices show TSPAN5 in the synaptosomal fraction. PSD-95, NLG1, and Synaptophysin are used as post- and presynaptic proteins, respectively. Tubulin is used as loading control. P, pellet; S, supernatant; Syn, synaptosomes; Tot H, total homogenate. Exact values are shown in . See also Figure S1 . Graphs represent mean ± SEM.

Article Snippet: α-VGlut1 Rabbit Polyclonal , Synaptic Systems , Cat. No. 135 303.

Techniques: Cell Culture, Immunolabeling, Purification

Journal: Cell Reports

Article Title: TSPAN5 Enriched Microdomains Provide a Platform for Dendritic Spine Maturation through Neuroligin-1 Clustering

doi: 10.1016/j.celrep.2019.09.051

Figure Lengend Snippet: TSPAN5 Depletion Does Not Affect Excitatory Synapse Function (A) Confocal images of DIV 18 rat hippocampal cultured neurons transfected at DIV 5 with scrambled, Sh-TSPAN5, or rescue constructs all co-expressing GFP and immunolabeled for PSD-95 (red) and VGluT1 (blue). Insets show higher magnification of dendrites highlighted in white boxes and colocalizing puncta between PSD-95 and VGluT1 (cluster, red). (B) Quantification relative to (A) of PSD-95-VGluT1 colocalizing puncta in GFP positive areas (number of puncta), puncta density (puncta/square micrometer) and average size (square micrometers). Gray charts show percentage of puncta distributed among the dendritic spines (white), the dendritic shaft (light grey), and the soma (dark grey). Colored pie charts show the distribution of PSD-95-VGlut1 colocalizing puncta on different subtypes of dendritic spines (stubby, thin, and mushroom). n = 16 neurons/condition. (C) Representative traces of AMPAR-mediated mEPSCs recorded from DIV 18 rat hippocampal cultured neurons transfected at DIV 5 with scrambled, Sh-TSPAN5, or rescue constructs. Scrambled, n = 24; Sh-TSPAN5, n = 25; rescue, n = 19. (D) Quantification of amplitude, frequency, area, and decay time of AMPAR-mediated mEPSCs relative to (C). Exact values are shown in . See also Figure S2 . Graphs represent mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus scrambled; °p < 0.05, °p < 0.01, and °p < 0.001 versus Sh-TSPAN5.

Article Snippet: α-VGlut1 Rabbit Polyclonal , Synaptic Systems , Cat. No. 135 303.

Techniques: Cell Culture, Transfection, Construct, Expressing, Immunolabeling

Journal: Cell Reports

Article Title: TSPAN5 Enriched Microdomains Provide a Platform for Dendritic Spine Maturation through Neuroligin-1 Clustering

doi: 10.1016/j.celrep.2019.09.051

Figure Lengend Snippet:

Article Snippet: α-VGlut1 Rabbit Polyclonal , Synaptic Systems , Cat. No. 135 303.

Techniques: Transduction, Recombinant, Modification, Software, Imaging

Journal: The Journal of Neuroscience

Article Title: Enhanced Excitatory Connectivity and Disturbed Sound Processing in the Auditory Brainstem of Fragile X Mice

doi: 10.1523/JNEUROSCI.2310-16.2017

Figure Lengend Snippet: Increased vGluT1/2 immunostained area in Fmr1 KO LSO neurons. A–D, Micrographs of WT and Fmr1 KO LSO sections immunostained against vGluT1 (green) and vGluT2 (red) at P20. E–H, Quantification of vGluT1/2 signal intensities (E, F) and immunolabeled area (G, H) of LSO coronal sections on P8, P20, and P60 WT and Fmr1 KO mice. Data are mean ± SEM. Scale bars: 20 μm. ***p < 0.001 (two-way ANOVA followed by Bonferroni post tests).

Article Snippet: Slices were subsequently incubated overnight in the primary antibody sera containing, depending on the experiment, rabbit α-FMRP (Abcam; dilution 1:500, RRID:AB_2278530), rabbit α-vGluT1 (Synaptic Systems; dilution 1:1000, RRID:AB_887875), guinea-pig α-vGluT2 (Synaptic Systems; dilution 1:1000, RRID:AB_887884), rabbit α-GluR1 (Millipore; dilution 1:1000, RRID:AB_2113602), chicken α-MAP2 (Neuromics; dilution 1:1000, RRID:AB_2314763), 3% normal donkey serum, and 0.2% Triton X-100, in 0.1 m PBS.

Techniques: Immunolabeling

Journal: The Journal of Neuroscience

Article Title: Enhanced Excitatory Connectivity and Disturbed Sound Processing in the Auditory Brainstem of Fragile X Mice

doi: 10.1523/JNEUROSCI.2310-16.2017

Figure Lengend Snippet: Increased vGluT1/2 immunostained area in Fmr1 KO LSO neurons. A–D, Micrographs of WT and Fmr1 KO LSO sections immunostained against vGluT1 (green) and vGluT2 (red) at P20. E–H, Quantification of vGluT1/2 signal intensities (E, F) and immunolabeled area (G, H) of LSO coronal sections on P8, P20, and P60 WT and Fmr1 KO mice. Data are mean ± SEM. Scale bars: 20 μm. ***p < 0.001 (two-way ANOVA followed by Bonferroni post tests).

Article Snippet: Slices were subsequently incubated overnight in the primary antibody sera containing, depending on the experiment, rabbit α-FMRP (Abcam; dilution 1:500, RRID:AB_2278530), rabbit α-vGluT1 (Synaptic Systems; dilution 1:1000, RRID:AB_887875), guinea-pig α-vGluT2 (Synaptic Systems; dilution 1:1000, RRID:AB_887884), rabbit α-GluR1 (Millipore; dilution 1:1000, RRID:AB_2113602), chicken α-MAP2 (Neuromics; dilution 1:1000, RRID:AB_2314763), 3% normal donkey serum, and 0.2% Triton X-100, in 0.1 m PBS.

Techniques: Immunolabeling